1、然后以3000r/min离心3min后进行检测。5. 超声波浸提法mL具塞试管中,用提取液定容至刻度, 然后将超声波清洗机的温度调为50,加热40min然后以3000r/min离心3min后进行检测。计算方法:取待测液,于663 nm与645 nm处测定吸光度值,按照Arnon公式计算叶绿素a和叶绿素b及叶绿素的含量 叶绿素a提取量(mgg )=12.7A6632.69A645 V (1000W) 叶绿素b提取量(mgg)= 22.9A645-4.68 A663V (1000叶绿素总提取量(mg )=叶绿素a提取量+叶绿素b提取量 式中:A663波长663 nm处的吸光度 A645波长645
2、nm处的吸光度 V提取液的体积(mL) W一 叶片质量(g)1. Arnon DL Copper enzymes in isolated chloioplastspolyphenol oxidase in Beta vulgarisJPlant Physiology,1949,24:1-5实验数据记录:直接浸提法 53h 3月20日3月22日波长A645A663绿色0.5510.5250.8160.5100.7560.476平均值0.7080.504红色0.2990.2470.3030.2490.3220.2530.3080.250老叶:叶绿素a提取量(mgg )= 0.389叶绿素b提取量(
3、mgg)= 0.167叶绿素总提取量(mg )=0.556新叶:叶绿素a提取量(mgg )= 0.195叶绿素b提取量(mgg)=0.063叶绿素总提取量(mg )=0.258加热浸提法 3月23日0.3250.9980.3330.9260.3820.8930.3470.9390.1170.3650.1270.3860.1150.3620.1200.371叶绿素a提取量(mgg )=0.727叶绿素b提取量(mgg)=0.237叶绿素总提取量(mg )=0.964叶绿素a提取量(mgg )=0.293叶绿素b提取量(mgg)=0.529叶绿素总提取量(mg )=0.822研磨法 3ml 3月2
4、5日0.4170.7220.6430.9680.5180.8220.5260.8370.5700.9860.5330.7630.3750.5680.4930.772叶绿素a提取量(mgg )=0.369叶绿素b提取量(mgg)=0.325叶绿素总提取量(mg )=0.694叶绿素a提取量(mgg )=0.340叶绿素b提取量(mgg)=0.307叶绿素总提取量(mg )=0.647超声波浸提法40min 3月27日0.2720.7400.3930.9870.2970.6500.3210.7920.1230.3400.1830.5300.1440.406叶绿素a提取量(mgg )=0.613叶绿
5、素b提取量(mgg)=0.226叶绿素总提取量(mg )=0.839叶绿素a提取量(mgg )=0.318叶绿素b提取量(mgg)=0.093叶绿素总提取量(mg )=0.411超声波研磨法 20min 3月27日0.8030.5110.6360.6220.3910.6870.4290.3970.2630.5380.5490.2750.5670.359叶绿素a提取量(mgg )=0.326叶绿素b提取量(mgg)=0.181叶绿素总提取量(mg )=0.507叶绿素a提取量(mgg )=0.273叶绿素b提取量(mgg)=0.148叶绿素总提取量(mg )=0.421超声波研磨法 40min
6、3月28日0.3270.2810.7550.4220.4660.3010.5160.3350.3660.5540.3630.6400.4070.5770.379叶绿素a提取量(mgg )=0.256叶绿素b提取量(mgg)=0.354叶绿素总提取量(mg )=0.610叶绿素a提取量(mgg )=0.290叶绿素b提取量(mgg)=0.146叶绿素总提取量(mg )=0.436超声波研磨法 60min 3月29日0.3410.8980.3390.8840.3290.8420.3360.8750.2900.7110.2190.5290.3150.7530.664叶绿素a提取量(mgg )=0.6
7、81叶绿素b提取量(mgg)=0.240叶绿素总提取量(mg )=0.921叶绿素a提取量(mgg )=0.513叶绿素b提取量(mgg)=0.213叶绿素总提取量(mg )=0.726超声波研磨法 70min 3月30日0.3680.9640.2620.3380.8820.3230.8390.1410.3530.1780.4860.1900.4720.1700.437叶绿素a提取量(mgg )=0.652叶绿素b提取量(mgg)=0.231叶绿素总提取量(mg )=0.883叶绿素b提取量(mgg)=0.123叶绿素总提取量(mg )=0.463超声波研磨法 80min 3月30日0.314
8、0.8210.8500.2940.7260.7990.1560.3950.1630.1460.3800.155叶绿素a提取量(mgg )=0.617叶绿素b提取量(mgg)=0.259叶绿素总提取量(mg )=0.876叶绿素a提取量(mgg )=0.308叶绿素b提取量(mgg)=0.113数据分析:由上表可知超声波研磨法的超声最佳时间是60min,并且加热浸提法的提取叶绿素含量较高。注明:红线部分是没有进行稀释的,而黑色的是稀释了10ml的,并且研磨法是稀释了3ml。因为红叶石楠的叶绿素没有人做过,所以我的提取剂以及配比是从有关植物叶绿素的提取的论文里参考的。参考文献:80%丙酮:1 油菜叶绿素提取方法的研究液料比1g:20ml青草叶绿素提取工艺的研究提取剂用量20 mL冬青叶叶绿素提取工艺研究研磨5min叶绿素提取 分离及观察体会
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